Production of Innervated Skeletal Muscle Fibers Using Human Induced Pluripotent Stem Cells

Only a limited number of large-scale protocols describe the production of mature skeletal muscle fibers from human induced pluripotent stem cells (hiPSCs). Here we describe a novel procedure for simultaneous differentiation of hiPSC into muscle cells and motor neurons, that generates innervated and contractile multinucleated skeletal muscle fibers with sarcomeric organization. Our protocol permits the production of expandable skeletal muscle progenitor cells and mature skeletal muscle fibers that can be used for the exploration of skeletal muscle differentiation for basic research, disease modeling, and drug discovery.